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TUNEL Assay Kit - BrdU-Red (ab66110)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (7)Q&A (7)References (145)

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In Situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit (ab66110)
  • Fluorescence Microscopy - In situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit (ab66110)
  • Immunohistochemistry - ab66110

Key features and details

  • Assay type: Quantitative
  • Detection method: Fluorescent
  • Platform: Flow cytometer, Fluorescence microscope
  • Assay time: 3 hr
  • Sample type: Adherent cells, Suspension cells, Tissue

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関連製品

医薬用外毒物

製品の概要

  • 製品名

    TUNEL Assay Kit - BrdU-Red
    DNA fragmentation キット 製品一覧
  • 検出方法

    Fluorescent
  • サンプルの種類

    Tissue, Adherent cells, Suspension cells
  • アッセイタイプ

    Quantitative
  • 全工程の試験時間

    3h 00m
  • 種交差性

    交差種: Mammals, Other species
  • 製品の概要

    TUNEL Assay Kit - BrdU-Red ab66110 uses a convenient and sensitive method to detect DNA fragmentation by flow cytometry and fluorescence microscopy in live cells.


    The TUNEL assay is used to detect DNA fragmentation, such as in apoptosis. It uses terminal deoxynucleotidyl transferase (TdT) to catalyze the incorporation of deoxynucleotides at the free 3'-hydroxyl ends of fragmented DNA. The deoxynucleotides are then labeled in a variety of ways for detection of the degree of DNA fragmentation.


    This TUNEL assay protocol is based on Br-dUTP (bromolated deoxyuridine triphosphate nucleotide), which can be more readily incorporated into DNA strand breaks by the TdT enzyme than other dUTP labels such as FITC, biotin or dioxigenin. The greater incorporation rate produces a brighter signal when the Br-dUTP sites are detected with an anti-BrdU monoclonal antibody directly labeled with a red fluorochrome.


    The BrdU-Red signal can be analyzed at Ex/Em 488/576 nm, with an optional 7-AAD counterstain at Ex/Em 488/655nm.


    This TUNEL assay kit includes both negative and positive control cells. It is designed to be suitable for studying DNA fragmentation in GFP-transfected cells.


    Tunel assay protocol summary:
    - fix cells / tissues with formaldehyde, or deparaffinize and rehydrate if paraffin sections, and wash
    - incubate cells in 70% ethanol for 30 min at 4ºC, or if tissues incubate with proteinase K solution for 5 min at room temp and refix with formaldehyde
    - wash
    - incubate in DNA labeling solution for 60 min at 37ºC
    - wash
    - incubate in antibody solution for 30 min at room temp
    - add 7-AAD / RNase A solution and incubate for 30 min at room temp
    - analyze with flow cytometry or fluorescent microscopy

  • 特記事項

    This product is manufactured by BioVision, an Abcam company and was previously called K404 ApoBrdU Red DNA Fragmentation Kit. K404-60 is the same size as the 60 test size of ab66110.

    This kit is BrdU-Red labeled (Ex/Em = 488/576 nm). It was previously called TUNEL Assay Kit - In situ BrdU-Red DNA Fragmentation.

    To use FITC (Ex/Em = 495/519 nm) as a label, we recommend TUNEL Assay Kit - FITC (ab66108).

    For chromogenic TUNEL staining, we recommend TUNEL Assay Kit - HRP-DAB ab206386.

    Find out more about the TUNEL method in the TUNEL staining / TUNEL assay guide.

  • 試験プラットフォーム

    Flow cytometer, Fluorescence microscope

製品の特性

  • 保存方法

    Please refer to protocols.
  • 内容 60 tests
    7-AAD/RNase Staining Buffer 1 x 30ml
    Anti-BrdU-Red Antibody 1 x 300µl
    Br-dUTP 1 x 480µl
    Negative Control Cells 1 x 5ml
    Positive Control Cells 1 x 5ml
    Reaction Buffer 1 x 600µl
    Rinse Buffer 1 x 120ml
    TdT Enzymes 1 x 45µl
    Wash Buffer 1 x 120ml
  • 研究分野

    • Kits/ Lysates/ Other
    • Kits
    • Apoptosis Kits
    • DNA Fragmentation
    • TUNEL Assay Kits
    • Kits/ Lysates/ Other
    • Kits
    • Molecular Biology Kits
    • DNA Isolation Kits
    • Kits/ Lysates/ Other
    • Kits
    • Apoptosis Kits
    • DNA Fragmentation
    • DNA ladder
    • Kits/ Lysates/ Other
    • Kits
    • Cell Damage Kits
    • Cell Damage
  • 関連性

    Internucleosomal DNA fragmentation is a hallmark of apoptosis in mammalian cells.

関連製品

  • Assay kits

    • TUNEL Assay Kit - HRP-DAB (ab206386)
    • TUNEL Assay Kit - FITC (ab66108)
  • Related Products

    • BrdU (5-bromo-2'-deoxyuridine), Thymidine analog (ab142567)

画像

  • In Situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit (ab66110)
    In Situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit (ab66110)

    Detection of DNA fragmention (TUNEL staining) using the negative and positive control cells (HL-60 untreated and treated with camptothecin). Cells were stained following the assay protocol. The fluorescence signal was detected and analyzed using BD FACScan System (Becton Dickinson).

  • Fluorescence Microscopy - In situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit (ab66110)
    Fluorescence Microscopy - In situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit (ab66110)This image is courtesy of an Abreview submitted by James Gahan

    TUNEL staining in whole mount Hydractinia echinata using In situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit (ab66110).

    Animals were fixed in 4% PFA in PBS for 1 hour and processed as per the protocol without proteinaseK treatment. In place of proteinaseK animals were permeabilised in 3% Triton in PBS for 15 minutes. Animals were counter-stained with DAPI.

  • Immunohistochemistry - ab66110
    Immunohistochemistry - ab66110Image from Hopkins J et al., PLoS Genet 10(7), fig 1C. Doi: 10.1371/journal.pgen.1004413. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Immunohistochemical analysis of paraffin embedded 5 micron thick testis tissues of 8 week old Stag3+/− and Stag3−/− (Stromal antigen) mice . Apoptotic cells were detected using In situ BrdU-Red DNA Fragmentation (TUNEL) Assay Kit (ab66110). DAPI was used as a counterstain.

プロトコール

  • Protocol Booklet

Click here to view the general protocols

データシートおよび資料

  • SDS download

  • Datasheet download

    Download

参考文献 (145)

ab66110 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab66110 は 145 報の論文で使用されています。

  • Najjar RS  et al. Raspberry polyphenols target molecular pathways of heart failure. J Nutr Biochem 124:109535 (2024). PubMed: 37984734
  • Han Y  et al. Extracellular vesicle-encapsulated CC16 as novel nanotherapeutics for treatment of acute lung injury. Mol Ther 31:1346-1364 (2023). PubMed: 36635966
  • Shi SX  et al. CD4+ T cells aggravate hemorrhagic brain injury. Sci Adv 9:eabq0712 (2023). PubMed: 37285421
  • Petsalaki E  et al. The abscission checkpoint senses chromatin bridges through Top2α recruitment to DNA knots. J Cell Biol 222:N/A (2023). PubMed: 37638884
  • Lee SY  et al. SH003 Causes ER Stress-mediated Apoptosis of Breast Cancer Cells via Intracellular ROS Production. Cancer Genomics Proteomics 20:88-116 (2023). PubMed: 36581346
View all Publications for this product

レビューと Q&A

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1-7 of 7 Abreviews

TUNEL staining on Lewis Lung Carcinoma cells in vitro

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
Worked very well on mouse Lewis Lung Carcinoma cells grown in vitro.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Louise Reynolds

Verified customer

投稿 Apr 05 2019

Excellent on human tumor tissue

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
This kit is very good when I used on human tumor sections. Just follow the manual and you will get good results. Also, if you want do double fluorescent staining for other antigen, you can use heat induced antigen retrieve too.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Hongwei Shao

Verified customer

投稿 Dec 19 2018

Tunel assay for cell death in E3 neural tube after Hhip electroporatin

Average Below average 2/5 (Ease of Use)
Abreviews
Abreviews
abreview image
It works on quail tissue.
It is a long procedure, it fades relatively fast.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Nov 28 2018

Flow Cyt

Average Average 3/5 (Ease of Use)
Abreviews
Abreviews
I tried to used (ab66110) For the Flow Cyt after fixing the cells it dose not give any differences between the positive and negative samples (dose not work)
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Nov 15 2018

Poor performance on cultured cells, average for tissue staining

Average Good 4/5 (Ease of Use)
Abreviews
Abreviews
abreview image
I purchased this kit with the purpose of staining cultured cells. It totally failed in that purpose and I would recommend the xxxx kit if your aim is to investigate apoptosis in cultured cells.
The protocol provided by Abcam does not mention any suggestion for cultured cells.
Additionally, the protocol is not well written. This includes a mistake on the manual with a recommended amount to pipette. You will easily find the mistake but this should not happen. I sent an email to Abcam and I hope they correct the mistake.

On the other hand, the kit works pretty well for tissues embedded in Paraffin. Attached is an example of mouse mammary gland. Magnification = 20X
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Sep 08 2017

Dual Caspase-3/TUNEL Fluorescence Staining

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
Frozen sections of snap frozen sekeletal muscle tissue with isopentane were used in the application. Sections were fixed in cold acetone for 10 minutes and then treated with 5% normal goat serum as a blocking reagent for 30 minutes.

*PBS was used for rinsing/washing solution in between steps.

No antigen retrieval used.

PART A: Caspase-3
i. Primary antibody ab52293 anti-Caspase-3 (1/200, overnight incubation at 4C)
ii. Secondary antibody ab7086 Goat anti-Rabbit IgG H&L FITC (1/200, 1 hour incubation at RT)

PART B: TUNEL
i. As per kit protocol from Part 2 (Detection by Fluorescence Microscopy), steps (a) to (l)
ii. After step (l), sections were dipped in DAPI diluted in PBS (1/10000) for 1 minute prior to final washing steps

Sections were coverslipped using fluoromount mountant and kept refridgerated at 4C in the dark until ready for visualisation.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Apr 07 2016

TUNEL staining in whole mount Hydractinia echinata

Good Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
Animals were fixed in 4% PFA in PBS from 1 hour and processed as per the protocol without proteinaseK treatment. In place of proteinaseK animals were permeabilised in 3%Triton in PBS for 15 minutes. Animals were counter-stained with DAPI.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. James Gahan

Verified customer

投稿 Oct 06 2014

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